本文選題：血管內(nèi)皮生長因子　切入點：腫瘤轉(zhuǎn)移相關(guān)基因MTA1　出處：《武漢大學(xué)》2012年博士論文

【摘要】：第一部分人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA表達(dá)的相關(guān)性研究 目的：研究人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與血管內(nèi)皮生長因子(VEGF)、增殖細(xì)胞核抗原(PCNA)表達(dá)的相關(guān)關(guān)系,并結(jié)合文獻(xiàn)探討其意義。 方法：采用免疫組化(SABC)和圖像分析技術(shù)檢測10例正常鼻咽部組織和75例人鼻咽癌標(biāo)本中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA表達(dá)水平,并分析表達(dá)的相關(guān)性。 結(jié)果：(1)腫瘤轉(zhuǎn)移相關(guān)基因MTA1、VEGF、PCNA在正常鼻咽部組織中的表達(dá)均為陰性；(2)腫瘤轉(zhuǎn)移相關(guān)基因MTA1、VEGF、PCNA在人鼻咽癌中均有陽性表達(dá),(3)人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA的表達(dá)水平呈明顯正相關(guān)(r=0.941和r=0.889,P0.05)。 結(jié)論：腫瘤轉(zhuǎn)移相關(guān)基因MTA1在人鼻咽癌的惡性生物學(xué)行為中可能起重要的作用,其可能成為人鼻咽癌治療的一個有效的靶點。 第二部分腫瘤轉(zhuǎn)移相關(guān)基因MTA1與鼻咽癌細(xì)胞浸潤和轉(zhuǎn)移的關(guān)系 目的：鼻咽癌具有很高的侵潤和轉(zhuǎn)移特性。而腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)是新近發(fā)現(xiàn)的一個腫瘤轉(zhuǎn)移候選基因。本研究通過比較MTA1基因在人鼻咽癌細(xì)胞高低轉(zhuǎn)移株的表達(dá)水平,探討MTA1表達(dá)與鼻咽癌細(xì)胞浸潤和轉(zhuǎn)移潛能的相關(guān)性。 方法：采用半定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)檢測增殖及轉(zhuǎn)移潛能不同的鼻咽癌細(xì)胞株6-10B和5-8F中MTA1的表達(dá)情況,用Boyden小室體外侵襲實驗檢測兩株細(xì)胞的體外轉(zhuǎn)移能力；用MTA1基因質(zhì)粒轉(zhuǎn)染低轉(zhuǎn)移潛能細(xì)胞株6-10B,通過半定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)檢測MTA1的表達(dá)；并檢測轉(zhuǎn)染前后細(xì)胞轉(zhuǎn)移能力的變化。 結(jié)果：MTA1在低轉(zhuǎn)移潛能細(xì)胞株6-10B中表達(dá)水平低,在高轉(zhuǎn)移潛能細(xì)胞株5-8F中表達(dá)水平高(p0.05)；體外侵襲實驗顯示高轉(zhuǎn)移株5-8F細(xì)胞體外侵襲力強(穿膜細(xì)胞相對百分率為46.3+2.4%)顯著高于低轉(zhuǎn)移潛能細(xì)胞株,其穿膜細(xì)胞相對百分率只有12.6+1.1%,轉(zhuǎn)染MTA1基因質(zhì)粒后,低轉(zhuǎn)移細(xì)胞株轉(zhuǎn)移潛能較未轉(zhuǎn)染細(xì)胞明顯增高(p0.05)。 結(jié)論：MTA1在人鼻咽癌細(xì)胞轉(zhuǎn)移過程中起重要作用,其作用機制以及作為腫瘤轉(zhuǎn)移治療靶點的可能性值得進(jìn)一步研究。 第三部分反義寡核苷酸對鼻咽癌細(xì)胞MTAl基因表達(dá)及侵襲性影響的研究 目的：合成人腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)的反義脫氧寡核苷酸,觀察其轉(zhuǎn)染后對人鼻咽癌CNE1細(xì)胞系中MTA1表達(dá)及鼻咽癌侵襲性的影響。 方法：免疫細(xì)胞染色觀察人工合成正義、反義及無意義MTA1基因片段轉(zhuǎn)染人鼻咽癌細(xì)胞CNE1后人腫瘤轉(zhuǎn)移相關(guān)基因的表達(dá),RT-PCR和Western blot檢測MTA1基因的(?)nRNA和蛋白質(zhì)的表達(dá)水平的變化。應(yīng)用Boyden小室體外侵襲力評價轉(zhuǎn)染前后細(xì)胞侵襲力的變化。 結(jié)果：反義寡核苷酸處理鼻咽癌細(xì)胞后,MTA1mRNA的表達(dá)下降(吸光度之比為24.09±0.22),與正義鏈組、無意義鏈組和空白對照組(34.23+0.15,33.22±0.28,35.18±0.22)相比差異有統(tǒng)計學(xué)意義,P0.05。Boyden小室體外侵襲實驗檢測顯示,反義寡核苷酸處理后鼻咽癌細(xì)胞的透膜能力明顯下降。 結(jié)論：MTA1同鼻咽癌的轉(zhuǎn)移能力密切相關(guān),反義寡核苷酸的轉(zhuǎn)染可阻遏鼻咽癌細(xì)胞中人腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)的表達(dá)。并因此有可能限制鼻咽癌的侵襲和轉(zhuǎn)移。
[Abstract]:The correlation of tumor metastasis related gene MTA1 and VEGF, PCNA expression in human nasopharyngeal carcinoma
Objective: To investigate the correlation between tumor metastasis related gene MTA1 and vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) expression in human nasopharyngeal carcinoma, and to explore its significance combined with literature.
Methods: immunohistochemistry and SABC were used to detect the expression levels of MTA1 and VEGF and PCNA in 10 normal nasopharyngeal tissues and 75 NPC cases, and the correlation between them was analyzed.
Results: (1) tumor metastasis related genes MTA1, VEGF, PCNA expression in normal nasopharyngeal tissues were negative; (2) tumor metastasis related genes MTA1, VEGF, PCNA were positive in human nasopharyngeal carcinoma, (3) human nasopharyngeal carcinoma metastasis related genes MTA1 and VEGF showed a positive correlation the expression level of PCNA (r=0.941, r=0.889, P0.05).
Conclusion: tumor metastasis related gene MTA1 may play an important role in malignant biological behavior of human nasopharyngeal carcinoma. It may become an effective target for treatment of NPC.
The relationship between the second part of tumor metastasis related gene MTA1 and the invasion and metastasis of nasopharyngeal carcinoma cells
Objective: nasopharyngeal carcinoma with invasion and metastasis is very high. And the characteristics of tumor metastasis associated gene (MTA1) is a recently discovered tumor metastasis genes. The expression level of MTA1 gene in human nasopharyngeal carcinoma cell lines with different metastatic cells, to investigate the correlation between MTA1 expression and the potential of invasion and metastasis of nasopharyngeal carcinoma.
Methods: using semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to detect the proliferation and metastatic potential of different nasopharyngeal carcinoma cell lines 6-10B and MTA1 in 5-8F expression, with Boyden chamber in vitro invasion assay of two cell lines in vitro metastasis; MTA1 gene transfected with low metastatic potential cell line 6-10B was detected by semi quantitative MTA1. Reverse transcriptase polymerase chain reaction; change detection and transfer ability before and after transfection.
Results: the MTA1 in low metastatic potential cell line 6-10B low expression in highly metastatic potential cell line 5-8F high expression level (P0.05); experiments show high metastatic 5-8F cells in vitro invasiveness in vitro invasion (transmembrane cell relative percentage of 46.3+2.4%) was significantly higher than that of low metastatic potential cell line, the cell membrane the relative percentage of only 12.6+1.1%, transfection of MTA1 gene plasmid, metastatic potential than non transfected cells was significantly higher in low metastatic cell line (P0.05).
Conclusion: MTA1 plays an important role in the metastasis of human nasopharyngeal carcinoma cells, and its mechanism and the potential as a therapeutic target for tumor metastasis are worth further research.
Study on the effect of third antisense oligonucleotides on MTAl gene expression and invasiveness in nasopharyngeal carcinoma cells
Objective: to synthesize antisense oligodeoxynucleotide of human tumor metastasis related gene (MTA1) and observe its effect on MTA1 expression and invasion of nasopharyngeal carcinoma cell line after transfection in human nasopharyngeal carcinoma CNE1 cell line.
Methods: To observe the effect of synthetic justice immune cells staining. The expression of antisense MTA1 gene transfection and significance of human nasopharyngeal carcinoma cells CNE1 human tumor metastasis related gene, detection of MTA1 gene RT-PCR and blot Western (?) expression of nRNA and protein. The application of Boyden chamber invasion force evaluation changes before and after transfection cell invasion.
Results: antisense oligonucleotide treatment of nasopharyngeal carcinoma cells, the expression of MTA1mRNA decreased (absorbance ratio of 24.09 + 0.22), and sense group, no significant chain group and blank control group (34.23+0.15,33.22 + 0.28,35.18 + 0.22) compared to the difference was statistically significant, P0.05.Boyden chamber in vitro invasion assay showed that antisense oligonucleotide treatment membrane ability of nasopharyngeal carcinoma cells decreased significantly.
Conclusion: MTA1 is closely related to the metastatic ability of nasopharyngeal carcinoma. The transfection of antisense oligonucleotides can inhibit the expression of human tumor metastasis related gene (MTA1) in nasopharyngeal carcinoma cells. Therefore, it may limit the invasion and metastasis of nasopharyngeal carcinoma.

【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R739.63

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